《芒果果實(shí)中有機(jī)酸分析方法高效液相色譜法》編制說明（征求意見稿）根據(jù)《廣西農(nóng)業(yè)農(nóng)村產(chǎn)業(yè)振興促進(jìn)會關(guān)于<芒果果實(shí)中有機(jī)酸測定高效液相色譜法>等6項團(tuán)體標(biāo)準(zhǔn)立項的通知》文件要求，本標(biāo)準(zhǔn)由廣西壯族自治區(qū)農(nóng)GB5009.157-2016食品安全國家標(biāo)準(zhǔn)食品NB/T12004-2016費(fèi)托合成水中有機(jī)酸、醇、醛、酮的NY/T2277-2012水果蔬菜中有機(jī)酸和陰離子的測定離子色500-2014煙草及煙草制品揮發(fā)性有機(jī)酸的測定氣相色譜-質(zhì)譜8657-2014苯乙烯-丁二烯生橡膠T/NAIA0200-2023葡萄酒中多種有機(jī)酸的測定T/NAIA0207-2023釀酒葡萄及葡萄酒中有機(jī)酸的測定液T/NAIA098-2021釀酒葡萄及葡萄酒中有機(jī)酸含量的測定的側(cè)定方法。適用于果汁及果汁飲料、碳酸飲料、固體飲料、膠基糖果、餅干、經(jīng)查閱,國內(nèi)外未制定有與利用高效液相色譜法分析芒果果實(shí)中有機(jī)酸的相本標(biāo)準(zhǔn)根據(jù)GB/T1.1—2020《標(biāo)準(zhǔn)化工作導(dǎo)則第1部分：標(biāo)準(zhǔn)化文件的結(jié)％）準(zhǔn)儲備液；a-酮戊二酸、富馬酸、沒食子酸標(biāo)準(zhǔn)品40.00mg用純凈水定容至10酸、檸檬酸、琥珀酸配置成濃度為800mg/L；草酸配置成濃度為400），13.19mg/kg、64.01～1541.38mg/kg、2.59～133.97mg/kg。檸檬酸、乳酸、圖1芒果果實(shí)中11種有機(jī)酸物質(zhì)含量4高效液相色譜法測定芒果中草酸、酒石酸、奎寧酸、蘋果酸、a-酮戊二在190～220nm之間的波長。這是因為有機(jī)酸（如乙酸、乳酸、檸檬酸等）通常在紫外光譜的遠(yuǎn)紫外區(qū)具有吸收峰。這一區(qū)域的吸收主要與有機(jī)酸的羧基（-COOH）基團(tuán)相關(guān)。190nm適合檢掃描波長范圍190～300nm，對草酸、酒石酸、奎寧酸、乳酸、乙酸、檸檬酸、富馬酸、琥珀酸、沒食子酸分別進(jìn)行光譜掃描。由圖2明Shim-packGISC18峰型偏圓，PerfectT3峰型較好，能將有機(jī)酸分離開來進(jìn)一步對流動相流速進(jìn)行優(yōu)化，選用0.01mol/LpH2.8磷酸二氫鉀中含3%行等度洗脫。結(jié)果表明流動相流速為0.5ml/min時，有機(jī)酸出峰效果最好（圖在進(jìn)行方法準(zhǔn)確度與精密度之前，首先對全國40份芒果樣品進(jìn)行檢測，發(fā)琥珀酸、沒食子酸含量最多582.40mg/kg14.83mg/kg、1731.58mg/250.14～571.40mg/kg，314.63～2161.58mg/kg，248.41～2211.27～2524.91mg/kg，0～120.95mg/kg，237.71～3187.08mg/kg，78.90~0～133.97mg/kg。設(shè)置25mg/kg、250mg/k計算回收率，確定方法準(zhǔn)確性及精密度，結(jié)果見表2。組分試樣組分濃度（mg/kg）理論加標(biāo)濃度（mg/kg）加標(biāo)試樣組分濃度（mg/kg）回收率(%)平均回收率(%)RSD(%)草酸2534.7298.2095.912.3033.5793.6034.1595.9225035.99103.28103.320.1836.05103.5235.96103.16500528.10103.5996.076.53469.5791.88473.8292.73酒石酸2542.29106.00105.810.6242.37106.3242.07250287.86108.83101.886.03262.7298.77260.9298.05500518.09100.46102.452.95545.00105.84521.05101.05奎寧酸2538.03102.44104.604.2039.78109.4437.90101.92250268.77102.54102.213.18259.6098.87275.44105.21500540.00105.51107.492.17561.52109.82548.15107.15蘋果酸50.522573.6692.5694.6374.7296.8074.1594.52250279.6298.8498.570.27296.2798.30296.9498.57500562.07102.31102.720.42566.22102.72a-酮戊二酸6.052530.2796.8893.922.9628.7990.9639.5393.92250257.86100.72100.312.45250.2597.68262.40102.54500546.55108.10107.540.55541.00106.99543.80107.55乳酸35.312562.25107.76105.572.0961.21103.6061.65105.36250305.86108.22106.702.85306.50108.48293.83103.41500570.31107.00104.262.58544.66101.87554.86103.9125.202550.0299.2897.3649.0695.4449.5497.36250270.7298.2198.690.448373.1599.18271.9298.69500536.25102.21103.70104.15548.80104.72檸檬酸195.8625221.84103.92104.363.16221.22101.44222.79107.72250461.31106.18105.440.74457.63104.71459.46105.44500706.81102.19102.130.07102.05706.56富馬酸2.612528.96105.400.7229.32106.84250263.52104.36103.520.84259.30102.68261.40103.52500507.40100.96102.01517.95103.07512.65102.01琥珀酸49.912574.1596.9698.2074.7799.4474.4698.20250293.4597.4296.640.89292.0096.84289.0895.67500501.0690.2392.882.33518.7193.76523.1094.64沒食子酸3.332528.2099.4899.770.4428.40100.2828.2299.56250260.33102.80101.93258.78102.18255.35100.81500527.05104.74103.890.84518.60103.05522.80103.89組分濃度（mg/L）峰面積RSD草酸80.005739975790345765150.44酒石酸400.004207684130684169180.92奎寧酸800.00494747508533501640蘋果酸800.005423725385485404600.35a-酮戊二酸3601823562533582170.55乳酸800.003227533188973208250.60800.003799283735803767540.84檸檬酸800.00694231675359684795富馬酸135205013230191337535琥珀酸800.00398647386644392645沒食子酸188081718248031852810稀釋梯度下有機(jī)酸的色譜圖，確定方法的校準(zhǔn)曲線及相關(guān)系數(shù)，結(jié)果見表3。序號組分線性范圍（mg/L）線性方程相關(guān)系數(shù)R21草酸3.125~400y=7710x-9540.60.99922酒石酸6.25~800y=1801.4x-1631.10.99933奎寧酸6.25~800y=191.08x+4791.50.99994蘋果酸6.25~800y=739.24x+290.790.99975a-酮戊二酸y=1796.25x-1585.60.99896乳酸6.25~800y=382.81x-509.860.999376.25~800y=509.53x-657.240.99958檸檬酸6.25~800y=811.76x+285.220.99909富馬酸y=10416x-1573.50.9997琥珀酸6.25~800y=516.21x-1050.50.9997沒食子酸y=12763x-4730.90.9997果果實(shí)中有機(jī)酸的平均回收率在92.88107.54％之間，相對標(biāo)準(zhǔn)偏差在0.076.53％之間，10種有機(jī)酸在液相色譜儀上響應(yīng)值的RSD為0.44%~酸檢出限為3.125mg/kg，定量限為6.25酸檢出限為3.125mg/kg，定量限為6.25的檢測方法處理樣品，計算回收率。試驗結(jié)果表明，25mg/kg、250mg組分(%)(%)RSD(%)草酸38.632561.7592.4894.522.0662.7296.3662.3194.72250270.5392.7695.953.00280.4396.72284.5398.36500533.0398.8899.96537.7099.81544.54101.18酒石酸33.602559.49103.56105.2960.08105.9260.20106.40250290.05102.58105.222.64296.00104.96303.87500525.3598.3594.233.81496.5592.59492.4091.76奎寧酸42.582565.7292.5695.432.6466.9197.3266.6896.40250313.98108.56102.155.59293.30100.29286.5897.60500554.44102.37105.393.52590.25109.53563.90104.26蘋果酸211.5325234.4291.5694.233.72236.0898.20234.7992.92250460.0799.42104.414.42471.75105.29482.83108.52500739.98105.69107.90761.28109.95751.86108.07a-酮戊二酸8.562531.4391.4892.934.0031.1090.1632.8597.16250275.62106.822.28266.78103.29264.32102.30500522.60102.81101.212.28519.91102.27501.3598.56乳酸20.682545.3298.5695.314.5044.9196.9243.2990.44250277.83102.86104.592.20288.68107.20279.99103.72500530.15101.89102.062.47518.8199.63543.97104.6636.492562.87105.5297.077.9559.0990.4060.3195.28250282.4498.3899.293.9995.86295.55103.62500544.47101.60104.472.39106.13564.91105.68檸檬酸30.4325106.7698.957.2853.5792.5654.8197.5225098.2899.23281.83100.56277.5998.86500101.74104.392.99548.38103.59569.57107.83富馬酸5.982530.8199.3298.634.6231.68102.8029.4293.76250266.67104.28104.27263.46102.99269.81105.53500547.93108.39104.573.27523.46103.50515.04101.81琥珀酸32.002557.86103.4495.6554.7190.8455.1792.68250277.9998.40102.093.92297.86106.34285.85101.54500571.97107.99107.940.61568.27107.25574.86108.54沒食子酸2538.1594.6095.844.7039.71100.8437.5292.08250252.8195.3295.454.91241.5990.83265.03100.21500535.76104.25102.432.04528.97102.89515.0組分試樣組分濃度（mg/kg）理論加標(biāo)濃度（mg/kg）加標(biāo)試樣組分濃度（mg/kg）回收率(%)平均回收率(%)RSD(%)草酸34.962561.67106.84104.344.0359.8399.4861.64106.72250291.26102.5299.183.92272.2494.91285.25500549.58102.92100.702.08537.02100.41528.7598.76酒石酸39.732564.4298.76100.2565.27102.1664.6999.84250295.37102.26100.49289.79100.02287.6999.18500540.13100.08100.430.66544.72101.00545.75101.20奎寧酸25.552550.2198.64100.645.4152.25106.8049.6796.48250269.0597.4096.27260.7294.07268.9897.37500528.75100.64100.290.39527.41100.37524.8499.86蘋果酸32.602559.19106.36105.9659.44107.3658.64104.16250283.21100.24100.07285.56101.18297.5798.79500532.0999.9099.820.40533.44100.17529.4999.38a-酮戊二酸6.742530.6595.6499.493.4632.31102.2831.88100.56250263.72102.79102.700.08263.33102.64263.41102.67500509.81100.61100.500.10509.13100.48508.74100.40乳酸2534.2598.0894.983.4234.8095.2833.8891.60250261.89100.36100.040.66259.2999.32261.50100.21500513.80100.5699.710.81505.4998.90508.9899.602538.7595.5693.352.5237.5890.8838.2693.60250277.24104.9596.427.81241.5890.69248.9493.63500532.90103.61102.480.71528.70102.77525.60102.15檸檬酸25.432550.77101.3697.923.3449.8297.5694.84250275.80100.1598.272.48264.2195.51273.3099.15500527.00100.31103.903.08550.30104.97557.60106.43富馬酸9.282534.83102.2098.993.3634.0899.2033.1795.5625099.94101.532.09103.94261.03100.70500509.0399.95100.000.04509.33100.01509.47100.04琥珀酸2545.97106.20101.285.4743.2495.2845.01102.36250273.88101.78101.39275.87102.58268.9599.81500514.2898.9799.670.61519.53100.02519.50100.02沒食子酸5.802528.4890.7299.448.3130.91100.4432.59107.16250252.7598.7899.620.73256.07255.6999.96500100.06100.000.06505.7799.99505.5299.94組分試樣組分濃度（mg/kg）理論加標(biāo)濃度（mg/kg）加標(biāo)試樣組分濃度（mg/kg）回收率(%)平均回收率(%)RSD(%)草酸42.722568.50105.252.3468.89104.6869.71107.96250294.95100.89103.562.73300.86103.26309.02106.52500553.68102.19102.720.47558.9103.13556.96102.85酒石酸80.4425103.8293.5297.357.90106.99106.20103.5292.32250340.72105.60343.79105.34348.83107.36500592.35102.38101.730.97591.51102.21583.38100.59奎寧酸33.942562.9796.414.4564.78101.3662.8893.76250298.86103.69102.830.89294.08101.86296.80102.94500550.50102.21103.76564.25104.96560.05蘋果酸23.212549.38104.68106.79107.6450.22108.04250262.3095.6495.65267.4097.68257.3093.64500523.86100.130.10524.19100.20523.21100.00a-酮戊二酸4.582530.3696.596.2328.4295.3627.4091.28250249.0297.7996.52242.7595.27245.8096.49500500.0299.0999.580.43503.9499.87503.5299.79乳酸2535.78101.88101.002.1935.97102.6434.9398.48250259.5499.6999.890.56261.61100.52258.9399.45500516.25101.19101.290.15516.36101.21517.63101.462543.8795.80103.316.4046.99108.2846.38105.84250278.70103.51103.560.18278.42103.40279.32103.76500515.5599.1399.340.28518.2099.66516.1399.24檸檬酸2570.55105.72106.0470.28104.6471.06107.76250306.20104.83103.780.97303.30103.67301.20102.83500545.50100.2899.81546.80100.54537.1098.60富馬酸8.702534.91104.84101.053.5633.86100.6497.68250257.9799.7198.40255.8798.87250.2396.61500508.2099.9098.910.99498.3397.93503.1998.90琥珀酸9.482532.9593.8896.765.1932.9493.84102.56250263.90101.77103.72269.78272.66105.27500519.40101.98101.600.41515.22101.15517.85101.67沒食子酸6.432530.2795.3695.370.2330.2295.1630.3395.60250261.62102.08102.050.56262.96102.61206.09101.46500506.66100.05100.220.17508.42100.40507.47100.21組分試樣組分濃度（mg/kg）理論加標(biāo)濃度（mg/kg）加標(biāo)試樣組分濃度（mg/kg）回收率(%)平均回收率(%)RSD(%)草酸47.772573.55102.8797.4874.77108.00250305.550.34306.52103.50304.77102.80500578.85106.22106.000.26576.16105.68578.22106.09酒石酸54.432580.75105.28104.1380.62104.7680.02102.36250307.51101.23100.10306.25100.73300.2698.33500551.0299.3298.940.44549.63199.04546.7998.47奎寧酸49.612593.233.7473.8697.0072.7592.56250301.21100.64100.950.31301.96100.94302.75101.26500549.81100.04100.630.55555.32553.17100.71蘋果酸77.7525104.77108.08106.912.27104.88108.52103.78250331.89101.66103.20337.80104.02337.52103.91500577.5299.9510